ObjectiveTo develop the Evidence Grading Scale for Ancient classical prescriptions in Traditional Chinese medicine， assess its reliability and validity， and apply it in practice to provide multi-source evidence for clinical practice guidelines development. MethodsLiterature retrieval was conducted to extract and screen existing evaluation dimensions， then the initial items were summarized using thematic analysis. Experts in the clinical medicine， medical history and literature participated in the Delphi questionnaire survey to evaluate and refine the items. An expert consensus meeting was conducted to finalize the included items， refine the method for items evaluation and evidence grading. The evidence quality rating scale for ancient classical traditional Chinese medicine （TCM） prescriptions was then established and tested for reliability and validity. ResultsThrough literature review， extraction， screening and summarization， a total of 3 dimensions and 12 initial items were formed. Questionnaires were sent to 69 experts to evaluate the initial items， with a questionnaire response rate of 100% and an expert authority coefficient of 0.92. All 12 items were retained for they had importance scores above 4. The Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine includes 3 dimensions with 12 items. The 3 dimensions includes ancient evidence， inheritance status， and modern application. Each dimension contains 4 items， and each item has a full score of 5 points. The evidence was rated as high-level， moderate-level， and low-level according to the final scores. The content validity index （CVI） of the 12 items was ＞0.9， the average CVI of the scale was 0.98， and the intraclass correlation coefficient （ICC） was 0.90. ConclusionThe Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine has good reliability and validity， which is practical for use in the development of TCM clinical guidelines and can better support clinical decision-making.

Cognitive dysfunction caused by blast traumatic brain injury (bTBI) is a serious neurological disease with high incidence, serious condition and poor prognosis. bTBI can lead to a series of symptoms such as short-term memory loss, inattention or multi-tasking difficulties. In severe cases, bTBI can develop into Alzheimer′s disease, which has a great impact on patients′ normal work and life. At present, researches on cognitive dysfunction caused by bTBI mainly involve model construction, pathogenesis, pathophysiological changes, diagnosis and treatment, etc., and the molecular mechanism of its occurrence remains to be further studied. Under normal physiological conditions, the release of excitatory and inhibitory neurotransmitters, the release and uptake of Ca 2+, oxidation and antioxidant systems, and the promotion and inhibition of apoptosis are in a dynamic balance. bTBI disturbs the balance, which will lead to the damage of nerve cells at the molecular level, thus resulting in the occurrence of cognitive dysfunction. To this end, the authors summarized the aspects of excitatory toxicity and Ca 2+homeostasis disorder, oxidative stress, inflammation and edema, apoptosis, etc., and reviewed the research progress on the molecular mechanism of cognitive dysfunction caused by bTBI, so as to provide a reference for the treatment and rehabilitation of cognitive dysfunction in patients with bTBI.

Objective To explore the pharmacodynamic substances of Simiao Wan for the treatment of hyperuricemia and gout.Methods The pharmacological model of hyperuricemia was established.The chemical components in vivo and in vitro of Simiao Wan were analyzed by UPLC-Q-Exactive-MS.Based on the components absorbed in blood,the"active ingredient-target-pathway"network of Simiao Wan for regulating hyperuricemia and gout was constructed by network pharmacology method.The key ingredients were used for molecular docking with key targets[uricogenase(XDH),uric acid transporter(ABCG2,GLUT9,OAT1,URAT1)and inflammatory targets(PTGS2,TLR2,TLR4)]of hyperuricemia and gout.Finally,experimental verification was conducted according to the results of molecular docking.Our aim is to identify the key pharmacodynamic substances of Simiao Wan for the treatment of hyperuricemia and gout.Results Eighty-nine components of Simiao Wan were identified by UPLC-Q-Exactive-MS analysis,including 74 components absorbed in blood,which were confirmed as candidate ingredients.Network pharmacology was used to constructed"components absorbed in blood-target-pathway"network,and components absorbed in blood were matched with 116 targets of hyperuricemia and 173 targets of gout.It is involved in the regulation of biological processes,such as glucose and lipid metabolism,oxidative stress,inflammatory response,ERK1 and ERK2 cascade,MAPK cascade,PI3K signal transduction.Moreover,Simiao Wan plays a role in regulating the network of hyperuricemia and gout through regulating blood lipids and atherosclerosis,apoptosis,AGE-RAGE,TNF,PI3K-Akt,MAPK,TLRs,JAK-STAT,NF-κB and other signaling pathways.Molecular docking studies showed that berberine,phellodendrine,magnoflorine,jatrorrhizine,palmatine,obacunone,limonin,atractylodin,taxifolin,atractylenolide Ⅲ and β-ecdysterone had good affinity with uric acid synthase,uric acid transporter and inflammatory targets.Western Blot test showed that taxifolin negatively regulates the expression of URAT1.The above-mentioned compounds were the main pharmacodynamic substances of Simiao Wan for the treatment of hyperuricemia and gout.Conclusion This article describes the main pharmacodynamic substances of Simiao Wan in the regulation of hyperuricemia and gout,which can provides a scientific basis for the clinical application,improvement in quality evaluation and standard of Simiao Wan.

Objective To investigate the effects of high-definition transcranial direct current stimulation(HD-tDCS)on the modulation of the H-reflex and M-wave during ankle dorsiflexion-plantar flexion fatigue tasks to provide direction for the application of HD-tDCS in mitigating neuromuscular fatigue.Methods Twenty healthy young male participants were recruited and randomly assigned to either the real stimulation or sham stimulation group,with 10 participants in each group.The intervention consisted of a 5-day single-blind HD-tDCS application(duration:20 min;intensity:2 mA;target:Cz).Baseline measurements of the H-reflex and M-wave under resting conditions,M-wave during maximal voluntary isometric contraction(MVIC)of the dorsiflexor muscle,and MVIC torque of the dorsiflexor and plantar flexor muscles were obtained.An ankle dorsiflexion fatigue task was performed to determine the time to achieve fatigue for the task.The same fatigue task was repeated and evaluated one day after the intervention.A repeated-measures two-factor(stimulation condition x pre/post fatigue)analysis of variance(ANOVA)was used to analyze the effects of independent variables on the mechanical properties of the muscles and α-motoneuron conduction characteristics.Results After fatigue,voluntary activation(VA),maximal H-reflex(Hmax),maximal M-wave(Mmax),and dorsiflexor and plantar flexor MVIC torques in both groups were significantly reduced compared with pre-fatigue levels(P＜0.05).However,compared to the real stimulation group,the sham stimulation group showed a more significant decline in VA and plantar flexor MVIC torque(P＜0.05).Conclusions A continuous 5-day HD-tDCS intervention can effectively increase α-motoneuron activity at the spinal segment.It can also exert an inhibitory effect on reducing information transmission capacity at the peripheral neuromuscular junction under the ankle dorsi-plantarflexion fatigue task.

Background and purpose:In 2016 the National Cancer Institute(NCI)decided stopping to use NCI-60 cell lines for drug screening,suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research.The reason for NCI-60 cells'retirement'was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials.Since the major cancer behaviors,such as proliferation and metastasis,are fundamentally altered with long-term culture,the tumor cell lines are not representative of the characteristics of cancer in patients.Currently,scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past.Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research.Methods:Breast cancer tissues were collected in the Department of Breast Surgery,Affiliated Hospital of Guizhou Medical University.The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University(approval number:2022 ethics No.313),and the collection and use of tumor tissues complied with the Declaration of Helsinki.The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium.After the cells proliferated,the media were replaced with DEME medium.Cell line STR genotyping was done to determine cell-specific genetic markers and identification.Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors.Results:We created 6 primary breast cancer cell lines.The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features,clinical diagnosis,therapeutic regimens,clinical effectiveness and prognostic outcomes.The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines.Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines.Conclusion:We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.

Objective:To evaluate the safety and efficacy of intravenous tirofiban in stent-assisted embolization of acute ruptured intracranial aneurysms.Methods:A total of 286 patients with acute ruptured intracranial aneurysms who received stent-assisted embolization in Department of Neurosurgery, Linyi People's Hospital from January 2020 to September 2022 were enrolled. According to different preoperative antiplatelet regiments, they were divided into aspirin combined with double resistant group (preoperatively taking orally loading dose of aspirin and clopidogrel, n=167) and tirofiban group (intravenously injecting tirofiban, n=119). Propensity score matching (PSM) was used to adjust for potential differences in age, gender, Hunt-Hess grading, hypertension history, diabetes history, smoking history, aneurysm location, aneurysm neck, aneurysm body-neck ratio, and stent types; incidences of perioperative hemorrhagic and ischemic complications, and neurological recovery status at discharge (scores of modified Rankin scale [mRS]≤2 as good recovery) were compared between the two groups. Results:After 1:1 PSM, 96 patients were included in each group. No significant difference in incidence of hemorrhagic complications was noted between the double resistant group (2.1%) and tirofiban group (0.0%, P>0.05). No significant difference in incidence of ischemic complications was noted between the double resistant group (9.3%, including 8 with intraoperative thrombosis and 1 with postoperative infarction) and tirofiban group (7.2%, including 6 with intraoperative thrombosis and 1 with postoperative infarction, P>0.05). No significant difference in good recovery rate at discharge was noted between double resistant group (86.4%) and tirofiban group (90.6%, P>0.05). Conclusion:In stent-assisted embolization therapy for acute ruptured intracranial aneurysms, preoperative intravenous tirofiban enjoys the same safety and efficacy compared with preoperative oral loading dose of aspirin and clopidogrel.

The main component of the gas in the fish storage tank is hydrogen sulfide. Hydrogen sulfide poisoning is a common occupational chemical poisoning among fishermen in summer, and acute hydrogen sulfide poisoning can manifest as toxic encephalopathy. This paper analyzes a patient with delayed encephalopathy suspected of acute hydrogen sulfide poisoning. The patient was unconscious for 18 days after waking up for 5 days after acute hydrogen sulfide poisoning. After waking up again, there were symptoms such as decreased limb muscle strength, ataxia, swallowing, dysarthria, and the clinical characteristics were significantly different from those of delayed encephalopathy caused by acute carbon monoxide poisoning, such as decreased cognitive function and damage to extrapyramidal system.

The main component of the gas in the fish storage tank is hydrogen sulfide. Hydrogen sulfide poisoning is a common occupational chemical poisoning among fishermen in summer, and acute hydrogen sulfide poisoning can manifest as toxic encephalopathy. This paper analyzes a patient with delayed encephalopathy suspected of acute hydrogen sulfide poisoning. The patient was unconscious for 18 days after waking up for 5 days after acute hydrogen sulfide poisoning. After waking up again, there were symptoms such as decreased limb muscle strength, ataxia, swallowing, dysarthria, and the clinical characteristics were significantly different from those of delayed encephalopathy caused by acute carbon monoxide poisoning, such as decreased cognitive function and damage to extrapyramidal system.

Objective:To investigate the abnormalities of gray matter volume (GMV) and the synergistic changes in different cerebral regions in the first-episode and early-onset depression (EOD) patients.Methods:A total of 60 patients with untreated EOD (EOD group) and 64 healthy controls (control group) matched for age, gender, and education underwent high-resolution T 1WI MR scans. Voxel-based morphometry was used to calculate the cerebral GMV. The difference in GMV between the two groups was compared with the t-test. Different brain regions were selected as seeds for structural covariation network (SCN) analysis. Spearman correlation model was used to analyze the correlation between the GMV in different cerebral regions and illness duration as well as the scores of Hamilton rating scale for depression (HAMD) 17 items in EOD group. Results:Compared to control group, the EOD group had significantly increased GMV in the right orbitofrontal cortex, right dorsolateral prefrontal cortex, right inferior parietal lobule, right superior parietal lobule and bilateral precuneus ( P<0.05, corrected by FDR). Based on the right orbitofrontal cortex and dorsolateral prefrontal cortex as seed regions, structural covariance analysis revealed that abnormal cooperative brain regions in EOD group, mainly distributed in the bilateral frontal lobe, parietal lobe, occipital lobe, temporal lobe, paralimbic system and cerebellum ( P<0.05, corrected by FDR). In EOD group, significant negative correlations were observed between the GMV in the right orbitofrontal cortex ( r=-0.314, P=0.015), the left precuneus ( r=-0.283, P=0.029), and illness duration. Significant positive correlations were observed between the GMV in the right dorsolateral prefrontal cortex and the scores of anxiety/somatization factor of HAMD17 ( r=0.331, P=0.010), the left precuneus and weight factor of HAMD17 ( r=0.255, P=0.049), respectively. Conclusions:Abnormal GMV changes are observed in some regions of the prefrontal and parietal lobule in patients with untreated EOD, accompanied by extensive covariant brain regions and additional structural connectivity. In addition, the abnormal GMV changes in some regions are associated with clinical features. Part of the prefrontal and parietal lobule may be the biomarkers to objectively evaluate abnormal brain structure in depression patients in the early stage.

Objective:To investigate the effect and mechanism of Myosin X on the radiosensitivity of non-small cell lung cancer (NSCLC) cell line H1975 in vitro. Methods:Western blot was applied to detect the expression level of Myosin X expression. The H1975 cell line with stable knockout of Myosin X (KO group) and infected with control virus (NC group) were constucted by using CRISPR/Cas9 technique. The knockout efficiency was validated. The radiosensitivity of two cell lines was measured by colony formation assay and single-hit multi-target model. γ-H 2AX focus formation test and RNA sequencing (RNAseq) analysis were employed to identify the regulatory mechanism of the radiosensitivity of lung cancer cell lines mediated by Myosin X. Results:The expression level of Myosin X in the H1975 cells was significantly up-regulated than those in other NSCLC cell lines (all P<0.01). The lentiviral vector of Myosin X sgRNA-Lenti-CRISPR v2 was successfully constructed. After the puromycin screening, H1975 cell lines with complete knockout of Myosin X and control cell lines (NC group) were obtained. Colony formation assay demonstrated that compared with the NC group, the radiosensitivity in the KO group was significantly higher (The D 0 value was decreased from 1.28 Gy to 1.03 Gy, SF 2 decreased from 0.29 to 0.21, and the sensitization ratio was 1.24). The γ-H 2AX focus formation test showed that the number of damage focus formed at 1 h and 6 h after irradiation in the KO group was significantly larger than that in the NC group ( P<0.05. RNAseq analysis indicated that the expression level of ISLR in the KO group was significantly down-regulated than that IN the NC group ( P<0.05). Conclusion:Knockout of Myosin X can increase the radiosensitivity of H1975 cells probably by interfering the repair of DNA double-strand damage and down-regulating the expression level of ISLR.